Equal total protein amounts were resolved on SDS-PAGE gels and western blotted using anti-Akt, anti-JNK, anti-PAK1, or anti-GFP (Cell Signaling Technology, Inc., Danvers, MA) antibodies.
Cells were harvested and lysed as in the cell surface biotinylation experiments, and then equal protein amounts were resolved via SDS PAGE for Western blotting.
The general flow structure is analyzed first and the extent of the LES region and the amount of resolved fluctuations are discussed.
Microsomes containing various Sec61 mutants were normalized for total protein by BCA protein assay (Pierce) and equivalent amounts of total protein resolved by SDS-PAGE next to a serial dilution of canine rough microsomes (CRM) containing a known concentration of the Sec61 complex.
Though the fraction of nucleotides closely interacting with protein in capsids is difficult to measure experimentally, it might be estimated from the amounts of RNA resolved in crystallographic or EM structures.
Log phase cells (with or without prior treatment with CD44 siRNA and with or without stimulation using Hrg/HA as indicated in figure legends) were harvested, lysed in Western lysis buffer (1% Triton x-100 in 50 mM Tris, 150mM NaCl, 1mM EDTA, pH7.4 and containing protease/phosphatase inhibitors) and equal amounts of proteins resolved by 8% SDS-PAGE.
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Precipitation simulation is complicated as it is a conditional process and daily precipitation amount is poorly resolved by regional scale predictors (Wilby and Dawson, 2007).
Each sample (indicated amount of total proteins was resolved by SDS-PAGE.
Protein amounts were determined using the Pierce BCA protein assay (Thermo Fisher Scientific, Rockford, IL, USA) and equal amounts of proteins were resolved by SDS PAGE using 4 12% gels (NuPAGE, Invitrogen Life Technologies).
